Re: Osteocyte fixation

From:

Hazel Stevens <hstevens@bioeng.ucsd.edu>

Hi John,
My first thought was that you may not have many viable osteocytes after
several days in culture due to difficulties in keeping the cells
nourished in an organ culture type model. However this may not be the case
if it is trabecular. The best method i can think of is to cut frozen
sections of the biopsy and do an LDH (lactate dehydrogenase) stain. This
picks up leaky membrane cells (dying or dead) from intact membrane
(viable). I have never tried this technique with paraffin sections so I
don't know if it works with those but maybe someone else has.
Hazel
Hazel Stevens,
Tissue Engineering Science Laboratory
6405 EBU1
Department of Bioengineering
University of California, San Diego,
9500 Gilman Drive,
La Jolla,
CA 92093-0412,USA
Tel: (858) 534 1765
Fax: (858) 822 0240
On Mon, 12 Feb 2001, john baker wrote:
> This is for one of our graduate students here in the Orthopaedic Research Lab.
>
> What is a good fixative to preserve osteocyte morphology in bone biopsies?
> The biopsy is from regenerated bone filling in a defect in a rat femoral
> cortex. The biopsy is maintained in tissue culture for several days then
> embedded in paraffin, sectioned, and stained with Alcian blue Hematoxylin.
> The purpose of the study is to acertain the viability of the osteocytes.
> If you know of a good way to demostrate cell viability within tissue please
> include that too. If you require more information to help answer this
> please let me know.
> Thank you, John for Ed Hoffler
>
> John A. Baker
> The University of Michigan
> Orthopaedic Research Laboratories
> Histology Unit
> 400 North Ingalls, G161
> Ann Arbor, MI 48109-0486
> my office 734-936-1635
> lab office 734-763-9674
>
>
>